Charlier P, Buisson G, Dideberg O, Wierenga J, Keck W, Laible G, Hakenbeck R
Institut de Physique, Crystallographie, Université de Liège, Belgium.
J Mol Biol. 1993 Aug 5;232(3):1007-9. doi: 10.1006/jmbi.1993.1452.
A genetically engineered water-soluble derivative of PBP2x of Streptococcus pneumoniae has been produced, purified and crystallized in a form suitable for X-ray diffraction analysis. The best crystals have been grown at 15 degrees C, from solutions containing 8% polyethylene glycol 10,000 at pH values ranging from 3.9 to 6.0. These crystals diffract to a resolution of 3.5 A and have a space group P6(1)22 (or enantiomorph) with unit cell dimensions of a = b = 162.2 A, c = 171.8 A, alpha = beta = 90 degrees, gamma = 120 degrees. The molecular mass and cell dimensions suggest that there is one molecule of enzyme per asymmetric unit. The breakdown of a chromogenic cephalosporin derivative diffused into a crystal reveals clearly that the enzyme is active in the crystalline state.
已制备、纯化并结晶出一种基因工程改造的肺炎链球菌PBP2x水溶性衍生物，其结晶形式适用于X射线衍射分析。最佳晶体是在15℃下，从pH值为3.9至6.0、含有8%聚乙二醇10000的溶液中生长得到的。这些晶体的衍射分辨率为3.5 Å，空间群为P6(1)22（或对映体），晶胞参数为a = b = 162.2 Å，c = 171.8 Å，α = β = 90°，γ = 120°。分子量和晶胞尺寸表明每个不对称单元中有一个酶分子。扩散到晶体中的一种显色头孢菌素衍生物的分解清楚地表明该酶在结晶状态下具有活性。
